Development of rapid PCR method for simultaneous identification of species, specific capsular type, and toxigenicity of Pasteurella sp. isolates

A molecular multiplex PCR method for identification of East European green frog species (Pelophylax ridibundus, P. cf. bedriagae and P. lessonae) and their hybrids was developed. This simple and rapid method can be used for identification of species-specific mitochondrial and nuclear DNA. The method is based on species-specific differences in primary structure of the subunit 1 of the mitochondrial cytochrome C oxidase gene (COI) and the intron-1 of the nuclear serum albumin gene (SAI-1). Based on the method, we analyzed numerous individuals of these species and their hybrids from East European Plain, the Crimea, the Caucasus, the Ural, as well as introduced populations from Western Siberia and the Kamchatka. In all cases, identification of species performed by use of the multiplex PCR method coincided with results of study of primary nucleotide sequences.

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A Simple and Reliable Single Tube Septuple PCR Assay for Simultaneous Identification of Seven Meat Species

Foods ◽

10.3390/foods10051083 ◽

2021 ◽

Vol 10 (5) ◽

pp. 1083

Author(s):

Zhendong Cai ◽

Song Zhou ◽

Qianqian Liu ◽

Hui Ma ◽

Xinyi Yuan ◽

...

Keyword(s):

Dna Sequences ◽

Animal Species ◽

Meat Product ◽

Meat Products ◽

Pcr Assay ◽

Specific Primers ◽

Meat Species ◽

Pcr Method ◽

Species Specific ◽

Simultaneous Identification

Multiplex PCR methods have been frequently used for authentication of meat product adulteration. Through screening of new species-specific primers designed based on the mitochondrial DNA sequences, a septuple PCR method is ultimately developed and optimized to simultaneously detect seven species including turkey (110 bp), goose (194 bp), pig (254 bp), sheep (329 bp), beef (473 bp), chicken (612 bp) and duck (718 bp) in one reaction. The proposed method has been validated to be specific, sensitive, robust and inexpensive. Taken together, the developed septuple PCR assay is reliable and efficient, not only to authenticate animal species in commercial meat products, but also easily feasible in a general laboratory without special infrastructures.

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Identification of Bacteroides forsythus in Subgingival Dental Plaque with the Aid of a Rapid PCR Method

Journal of Dental Research ◽

10.1177/00220345970760070701 ◽

1997 ◽

Vol 76 (7) ◽

pp. 1376-1380 ◽

Cited By ~ 38

Author(s):

J.H. Meurman ◽

J. Wahlfors ◽

A. Korhonen ◽

P. Alakuijala ◽

P. Väisänen ◽

...

Keyword(s):

Dental Plaque ◽

Subgingival Plaque ◽

Chain Reaction ◽

Rapid Pcr ◽

Microbiological Methods ◽

Pcr Method ◽

Polymerase Chain ◽

Pre Treatment ◽

Culture Positive ◽

Positive Results

Bacteroides forsythus has been shown to be prevalent among patients with periodontitis. Conventional microbiological methods used to identify this bacterium, however, are laborious and time-consuming and are therefore not well-suited for screening purposes. We have developed a polymerase chain-reaction (PCR) method which is rapid, specific, and simple to perform and does not require other sample pre-treatment except a brief centrifugation. This method was applied to the detection of B. forsythus in subgingival plaque of 58 periodontitis patients. When compared with the results of conventional culturing, the PCR method always confirmed the culture-positive results, while none of the PCR negative samples was shown to be culture-positive. The PCR method appeared to give more than double the number of samples positive for B. forsythus than culturing (89.7% vs. 37.9%). The analysis requires less than 4 hrs to perform, and is specific only to B. forsythus and sensitive enough to detect fewer than 5 bacteria.

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